The Effect of Direct-Fed Microbials on In-Vitro Rumen Fermentation of Grass or Maize Silage
Direct-fed microbial products (DFM) are probiotics that can be used advantageously in ruminant production. The in vitro gas production technique (IVGPT) is a method to simulate rumen fermentation and can be used to measure degradation, gas production, and products of fermentation of such additives....
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MDPI AG
2023-04-01
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| Series: | Fermentation |
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| Online Access: | https://www.mdpi.com/2311-5637/9/4/347 |
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| author | Rajan Dhakal Giuseppe Copani Bruno Ieda Cappellozza Nina Milora Hanne Helene Hansen |
| author_facet | Rajan Dhakal Giuseppe Copani Bruno Ieda Cappellozza Nina Milora Hanne Helene Hansen |
| author_sort | Rajan Dhakal |
| collection | DOAJ |
| description | Direct-fed microbial products (DFM) are probiotics that can be used advantageously in ruminant production. The in vitro gas production technique (IVGPT) is a method to simulate rumen fermentation and can be used to measure degradation, gas production, and products of fermentation of such additives. However, inter-laboratory differences have been reported. Therefore, tests using the same material were used to validate laboratory reproducibility. The objective of this study was to assess the effect of adding two DFM formulations on fermentation kinetics, methane (CH<sub>4</sub>) production, and feed degradation in two different basal feeds while validating a newly established IVGPT laboratory. Six treatments, with three replicates each, were tested simultaneously at the established IVGPT lab at the University of Copenhagen, and the new IVGPT lab at Chr. Hansen Laboratories. Maize silage (MS) and grass silage (GS) were fermented with and without the following DFM: P1: <i>Ligilactobacillus animalis</i> and <i>Propionibacterium freudenreichii</i> (total 1.5 × 10<sup>7</sup> CFU/mL), P2: P1 with added <i>Bacillus subtilis</i> and <i>B. licheniformis</i> (total 5.9 × 10<sup>7</sup> CFU/mL). The DFM were anaerobically incubated in rumen fluid and buffer with freeze-dried silage samples for 48 h. Total gas production (TGP: mL at Standard Temperature and Pressure/gram of organic matter), pH, organic matter degradability (dOM), CH<sub>4</sub>concentration (MC) and yield (MY), and volatile fatty acid (VFA) production and profiles were measured after fermentation. No significant differences between the laboratories were detected for any response variables. The dOM of MS (78.3%) was significantly less than GS (81.4%), regardless of the DFM added (P1 and P2). There were no significant differences between the effects of the DFM within the feed type. MS produced significantly more gas than GS after 48 h, but GS with DFM produced significantly more gas at 3 and 9 h and a similar gas volume at 12 h. Both DFM increased TGP significantly in GS at 48 h. There was no difference in total VFA production. However, GS with and without probiotics produced significantly more propionic acid and less butyric acid than MS with and without probiotics. Adding P2 numerically reduced the total methane yield by 4–6% in both MS and GS. The fermentation duration of 48 h, used to determine maximum potential dOM, may give misleading results. This study showed that it is possible to standardize the methodology to achieve reproducibility of IVGPT results. Furthermore, the results suggest that the P2 DFM may have the potential to reduce CH<sub>4</sub> production without affecting organic matter degradation. |
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| spelling | doaj.art-b1a746a55b664057bd940579cbf1dab12023-11-17T19:10:36ZengMDPI AGFermentation2311-56372023-04-019434710.3390/fermentation9040347The Effect of Direct-Fed Microbials on In-Vitro Rumen Fermentation of Grass or Maize SilageRajan Dhakal0Giuseppe Copani1Bruno Ieda Cappellozza2Nina Milora3Hanne Helene Hansen4Department of Veterinary and Animal Sciences, University of Copenhagen, Grønnegårdsvej 3, 1870 Frederiksberg C, DenmarkChr. Hansen Animal and Plant Health & Nutrition, Bøge Alle 10-12, 2970 Hørsholm, DenmarkChr. Hansen Animal and Plant Health & Nutrition, Bøge Alle 10-12, 2970 Hørsholm, DenmarkChr. Hansen Animal and Plant Health & Nutrition, Bøge Alle 10-12, 2970 Hørsholm, DenmarkDepartment of Veterinary and Animal Sciences, University of Copenhagen, Grønnegårdsvej 3, 1870 Frederiksberg C, DenmarkDirect-fed microbial products (DFM) are probiotics that can be used advantageously in ruminant production. The in vitro gas production technique (IVGPT) is a method to simulate rumen fermentation and can be used to measure degradation, gas production, and products of fermentation of such additives. However, inter-laboratory differences have been reported. Therefore, tests using the same material were used to validate laboratory reproducibility. The objective of this study was to assess the effect of adding two DFM formulations on fermentation kinetics, methane (CH<sub>4</sub>) production, and feed degradation in two different basal feeds while validating a newly established IVGPT laboratory. Six treatments, with three replicates each, were tested simultaneously at the established IVGPT lab at the University of Copenhagen, and the new IVGPT lab at Chr. Hansen Laboratories. Maize silage (MS) and grass silage (GS) were fermented with and without the following DFM: P1: <i>Ligilactobacillus animalis</i> and <i>Propionibacterium freudenreichii</i> (total 1.5 × 10<sup>7</sup> CFU/mL), P2: P1 with added <i>Bacillus subtilis</i> and <i>B. licheniformis</i> (total 5.9 × 10<sup>7</sup> CFU/mL). The DFM were anaerobically incubated in rumen fluid and buffer with freeze-dried silage samples for 48 h. Total gas production (TGP: mL at Standard Temperature and Pressure/gram of organic matter), pH, organic matter degradability (dOM), CH<sub>4</sub>concentration (MC) and yield (MY), and volatile fatty acid (VFA) production and profiles were measured after fermentation. No significant differences between the laboratories were detected for any response variables. The dOM of MS (78.3%) was significantly less than GS (81.4%), regardless of the DFM added (P1 and P2). There were no significant differences between the effects of the DFM within the feed type. MS produced significantly more gas than GS after 48 h, but GS with DFM produced significantly more gas at 3 and 9 h and a similar gas volume at 12 h. Both DFM increased TGP significantly in GS at 48 h. There was no difference in total VFA production. However, GS with and without probiotics produced significantly more propionic acid and less butyric acid than MS with and without probiotics. Adding P2 numerically reduced the total methane yield by 4–6% in both MS and GS. The fermentation duration of 48 h, used to determine maximum potential dOM, may give misleading results. This study showed that it is possible to standardize the methodology to achieve reproducibility of IVGPT results. Furthermore, the results suggest that the P2 DFM may have the potential to reduce CH<sub>4</sub> production without affecting organic matter degradation.https://www.mdpi.com/2311-5637/9/4/347methane (CH<sub>4</sub>)in-vitro fermentationdirect-fed microbesruminantsprobiotics<i>Lactobacillus</i> |
| spellingShingle | Rajan Dhakal Giuseppe Copani Bruno Ieda Cappellozza Nina Milora Hanne Helene Hansen The Effect of Direct-Fed Microbials on In-Vitro Rumen Fermentation of Grass or Maize Silage Fermentation methane (CH<sub>4</sub>) in-vitro fermentation direct-fed microbes ruminants probiotics <i>Lactobacillus</i> |
| title | The Effect of Direct-Fed Microbials on In-Vitro Rumen Fermentation of Grass or Maize Silage |
| title_full | The Effect of Direct-Fed Microbials on In-Vitro Rumen Fermentation of Grass or Maize Silage |
| title_fullStr | The Effect of Direct-Fed Microbials on In-Vitro Rumen Fermentation of Grass or Maize Silage |
| title_full_unstemmed | The Effect of Direct-Fed Microbials on In-Vitro Rumen Fermentation of Grass or Maize Silage |
| title_short | The Effect of Direct-Fed Microbials on In-Vitro Rumen Fermentation of Grass or Maize Silage |
| title_sort | effect of direct fed microbials on in vitro rumen fermentation of grass or maize silage |
| topic | methane (CH<sub>4</sub>) in-vitro fermentation direct-fed microbes ruminants probiotics <i>Lactobacillus</i> |
| url | https://www.mdpi.com/2311-5637/9/4/347 |
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